Reblastatin derivative and use thereof in neurological diseases

Abstract

Provided are a reblastatin derivative and use thereof in neurological diseases. Modifying the hydroxyl group at position 18 of reblastatin improves the therapeutic activity on the nervous system. In addition, especially when the hydroxyl group is replaced with a methyl group, the lipid solubility of the compound can be improved, thereby improving the brain penetration efficiency.

Description

A riburamycin derivative and its use in neurological diseases Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a riburamycin derivative and its use in nervous system diseases. Background Technology

[0002] Excitotoxicity is a common pathological phenomenon in many brain diseases, and it is widely present in diseases such as epilepsy, Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS). It is characterized by the large accumulation of excitatory neurotransmitters, mainly glutamate, which causes toxic damage to neurons and triggers a series of neuropathological changes, including: (1) causing abnormal discharge of neurons, which can induce epilepsy in extreme cases; (2) overactivating intracellular nucleases and proteolytic enzymes in neurons, causing neurons to lose their homeostasis; (3) causing oxidative stress damage to neurons; and (4) in severe cases, directly inducing neuronal apoptosis or autophagy.

[0003] Excitotoxicity plays a crucial role in the pathogenesis of various neurological diseases, and is considered a key factor in disrupting normal brain function and leading to neurodegenerative diseases. Previously, researchers primarily focused on the repair strategies neurons might employ after excitotoxic injury: alleviating or reversing the pathological phenotype of the disease through methods such as drug or molecular interventions and cell therapy. For example, the increasingly reported methods of neural stem cell transplantation and inhibition of neuronal apoptosis have brought hope for the treatment of diseases such as ALS, AD, and PD. However, these measures also have some limitations, such as low stem cell survival rates and difficulties in integrating transplanted neurons into existing neural networks during cell transplantation therapy.

[0004] One hypothesis is that the limited effectiveness of such treatments stems from their failure to stop the "attack mechanisms" that are present throughout the course of the disease. Addressing these upstream attack mechanisms might yield better therapeutic results.

[0005] Astrocytes maintain normal glutamate levels in the neural microenvironment through the uptake and release of glutamate, with the glutamate transporter 1 (GLT1; also known as excitatory amino acid transporter 2, EAAT2) playing the most important role in glutamate clearance. GLT1 is a membrane-integrated protein specifically expressed by astrocytes that actively transports glutamate from the extracellular to intracellular environment, maintaining glutamate concentration within the normal range in the neural microenvironment. Intracellular glutamate is converted to glutamine by glutamine synthetase (GS) and enters metabolic pathways. GLT1 can also prevent neuronal overexcitation by rapidly clearing glutamate from receptors on the postsynaptic membrane. Reports show that the concentration difference of glutamate between intracellular and extracellular environments can reach up to 10,000 times under the action of GLT1. Studies have confirmed that GLT1 is the most important inward glutamate transporter in the brain, responsible for nearly 80% of glutamate clearance.

[0006] Evidence suggests that GLT1 is abnormally downregulated in various neurological diseases, leading to a severe loss of glutamate clearance capacity in astrocytes, which may be a significant cause of abnormal glutamate accumulation. Studies on patients with refractory temporal lobe epilepsy have found that both GLT1 mRNA and protein are downregulated in the epileptogenic focus (sclerotic hippocampus). Alzheimer's disease (AD) and epilepsy exhibit some comorbidity; multiple studies have confirmed excitotoxic neuronal damage in the brains of AD patients, and significantly reduced GLT1 protein levels have been found in autopsy brain tissue from AD patients. Studies on autopsy brain tissue from ALS patients have found GLT1 protein loss of up to 90%, and subsequent functional studies indicate that abnormal internalization and protein degradation of GLT1 under pathological conditions may be important reasons for its insufficient protein levels. Furthermore, researchers have also observed GLT1 downregulation in mouse models of tuberous sclerosis.

[0007] Functional studies at the animal level have provided conclusive evidence for the relationship between GLT1 loss and disease phenotypes. First, GLT1-transgenic mice exhibit anti-seizure capabilities. Under the induction of epileptogenic drugs (pilocarpine, a muscarinic acetylcholine receptor agonist used to establish rodent models of epilepsy via intraperitoneal injection), the proportion of hippocampal neuronal death, the degree of moss fiber sprouting, and other hippocampal sclerosis-related pathological phenotypes in GLT1-transgenic mice are reduced, and the frequency of spontaneous chronic epileptic seizures is alleviated by approximately 50%. Conversely, inhibiting GLT1 expression by injecting antisense RNA leads to a large accumulation of glutamate in the mouse brain, inducing neurodegenerative diseases and progressive paralysis. These studies demonstrate that astrocytes play a crucial neuroprotective role through their specifically expressed GLT1 molecules, and the loss of GLT1 is sufficient to cause excitotoxic damage to the nervous system.

[0008] In summary, the abnormal downregulation of GLT1 protein may be an important reason for the excessive accumulation of glutamate in the brains of patients with epilepsy, AD, and ALS. In recent years, researchers have proposed the hypothesis of excessive GLT1 protein degradation, finding that HSP90 protein promotes 20S proteasome-dependent GLT1 protein degradation by recruiting GLT1 to the proteasome. On the other hand, reactive astrocyte proliferation is a common pathological phenomenon in almost all brain diseases, and this process itself is accompanied by increased expression of HSP90 molecules. Therefore, treatment with HSP90 inhibitors can increase GLT1 protein levels in brain diseases such as temporal lobe epilepsy and AD. Regarding therapeutic effects, in mice with kaempferol-induced chronic temporal lobe epilepsy, two HSP90 inhibitors, 17AAG and NVP-HSP990, can alleviate epileptic seizures; in an AD mouse model, NVP-HSP990 can reduce epileptiform discharges in the brain and alleviate cognitive decline. Interestingly, because the expression level of HSP90 in astrocytes is much lower than that in neurons, while the overall expression level of HSP90 in the brain is comparable to that in other tissues and organs, the dosage of HSP90 inhibitors used in the treatment of brain diseases is much lower than the severe doses used in anti-tumor therapy, yet they can still increase GLT1 protein levels and exert anti-epileptic and anti-AD effects. These research results indicate that HSP90 inhibitors can be effective in treating brain diseases with neuroexcitotoxicity, such as epilepsy and AD, even at low doses.

[0009] However, existing HSP90 inhibitors have certain problems. For example, both 17AAG and NVP-HSP990 have their own drawbacks: 17AAG has poor drug-like properties (low water solubility, low bioavailability, and toxicity); NVP-990 has been reported to have optic nerve toxicity in clinical trials; (2) more importantly, HSP90 inhibitors have significant limitations in treating brain diseases (such as permeability to the blood-brain barrier). Therefore, it is necessary to further optimize and modify them to target the treatment of brain diseases.

[0010] Reblastatin, a type of anilinexamycin, has the following structural formula:

[0011] Compared to 17AAG, this compound has a phenolic structure instead of a benzoquinone structure on the benzene ring, thus reducing its toxicity. However, it also suffers from extremely low brain-crossing efficiency, making it necessary to modify its structure to improve this efficiency. Summary of the Invention

[0012] The purpose of this invention is to provide a ribramycin derivative and its use in neurological diseases. By modifying the hydroxyl group at position 18 of ribramycin, its neurological therapeutic activity is enhanced. Furthermore, especially when replaced with a methyl group, the compound's lipid solubility is increased, thereby improving its brain penetration efficiency.

[0013] To achieve the above-mentioned objectives, the technical solution of this invention is as follows:

[0014] In a first aspect, the present invention provides the use of a ribramycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs in the preparation of a treatment for and / or prevention of neurological diseases.

[0015] The structural formula of the riburamycin derivative is shown in Formula I:

[0016] Wherein, R is selected from H; halogen; amino; substituted or unsubstituted C1-5 alkyl, C1-5 alkoxy, C3-10 cycloalkyl, C1-5 ester, aryl, amino, heterocyclic, and the substituted group is selected from at least one of C1-C5 alkyl, carbonyl, hydroxyl, C1-C5 alkyl, halogen, amino, aryl, heterocyclic.

[0017] Preferably, R is selected from substituted or unsubstituted C1-5 alkyl, C1-5 alkoxy, C3-10 cycloalkyl, ester, aryl, amino, heterocyclic groups, and the substituted substituent is selected from at least one of C1-C5 alkyl, carbonyl, hydroxyl, C1-C5 alkyl, halogen, amino, aryl, and heterocyclic groups.

[0018] More preferably, R is selected from substituted or unsubstituted C1-5 alkyl, C1-5 ester, or heterocyclic groups, and the substituted substituent is selected from at least one of carbonyl, hydroxyl, C1-C5 alkyl, halogen, amino, aryl, or heterocyclic groups; the heteroatom in the heterocyclic group is selected from at least one of N, S, B, or O.

[0019] More preferably, R is selected from substituted or unsubstituted C1-3 alkyl, C1-5 ester, or heterocyclic groups, and the substituted substituent is selected from at least one of C1-C5 alkyl, amino, or heterocyclic groups; the heteroatom in the heterocyclic group is selected from at least one of N, S, B, or O.

[0020] More preferably, R is selected from C1-3 alkyl, -OCO-Ra, substituted heterocyclic groups, and Ra and the substituted substituents are each independently selected from at least one of C1-C3 alkyl, amino, and heterocyclic groups; the heteroatom in the heterocyclic group is selected from at least one of N, S, B, and O.

[0021] More preferably, R is selected from C1-3 alkyl, -OCO-Ra, substituted heterocyclic groups, and Ra is selected from at least one of amino and heterocyclic groups; the substituted substituent is selected from C1-C3 alkyl, and the heteroatom in the heterocyclic group is selected from at least one of N, B, and O.

[0022] More preferably, R is selected from C1-3 alkyl, -OCO-Ra, at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B or O, and Ra is selected from at least one amino group or N-containing six-membered heterocyclic group.

[0023] More preferably, R is selected from methyl, -OCO-Ra, at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B and O, and Ra is selected from at least one C1-3 alkyl-substituted amino group, piperazine group, and C1-3 alkyl-substituted piperazine group.

[0024] More preferably, R is selected from methyl, -OCO-Ra, at least one methyl-substituted pentacyclic heterocyclic group containing B or O, and Ra is selected from at least one of dimethylamino, methylethylamino, diethylamino, piperazine, or methyl-substituted piperazine.

[0025] More preferably, the R is selected from methyl, -OCO-Ra, Ra is selected from dimethylamino,

[0026] Most preferably, R is a methyl group.

[0027] Preferably, the neurological disease is a brain disease.

[0028] Preferably, the neurological disease is selected from at least one of epilepsy, Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS).

[0029] Further preferred are at least one of epilepsy and Parkinson's disease.

[0030] Secondly, the present invention provides a riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs, with the structural formula shown in Formula I:

[0031] The R is selected from methyl, -OCO-Ra, or substituted heterocyclic groups, and the Ra and the substituted substituents are each independently selected from at least one of C1-C3 alkyl, amino, or heterocyclic groups; the heteroatom in the heterocyclic group is selected from at least one of N, S, B, or O.

[0032] Preferably, R is selected from C1-3 alkyl, -OCO-Ra, substituted heterocyclic groups, and Ra is selected from at least one of amino and heterocyclic groups; the substituted substituent is selected from C1-C3 alkyl, and the heteroatom in the heterocyclic group is selected from at least one of N, B, and O.

[0033] More preferably, R is selected from C1-3 alkyl, -OCO-Ra, at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B or O, and Ra is selected from at least one amino group or N-containing six-membered heterocyclic group.

[0034] More preferably, R is selected from methyl, -OCO-Ra, at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B and O, and Ra is selected from at least one C1-3 alkyl-substituted amino group, piperazine group, and C1-3 alkyl-substituted piperazine group.

[0035] More preferably, R is selected from methyl, -OCO-Ra, at least one methyl-substituted pentacyclic heterocyclic group containing B or O, and Ra is selected from at least one of dimethylamino, methylethylamino, diethylamino, piperazine, or methyl-substituted piperazine.

[0036] More preferably, the R is selected from methyl, -OCO-Ra, Ra is selected from dimethylamino,

[0037] Most preferably, R is a methyl group.

[0038] Thirdly, the present invention provides a method for preparing the above-mentioned libramycin derivative, selected from the following methods:

[0039] R has the same definition as described above.

[0040] Preferably, when R is a methyl or substituted heterocyclic group, the preparation method is as follows:

[0041] When R is -OCO-Ra, the preparation method is as follows:

[0042] Fourthly, the present invention provides a pharmaceutical composition comprising the aforementioned riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and pharmaceutically acceptable carriers or excipients.

[0043] The term "carrier" is well-known in the field and includes pharmaceutically acceptable materials, components, or delivery agents suitable for administration of the compounds of the present invention to mammals. Carriers include liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials that participate in carrying or delivering the subject matter from one part of an organ or body to another. Each carrier must be "acceptable" in terms of compatibility with other components of the formulation or harmlessness to the subject. Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium methylcellulose, ethylcellulose, and cellulose acetate; tannins, malt, gelatin, and talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; and glycols, such as propylene glycol and polyols, such as glycerol and sorbitol. Pear alcohol, mannitol, and polyethylene glycol, esters such as ethyl oleate and ethyl laurate, agar, buffers such as magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogens, isotonic saline, ethanol, phosphate buffer and other non-toxic compatibility substances used in pharmaceutical formulations, wetting agents, emulsifiers and lubricants such as sodium dodecyl sulfate and stearate, as well as colorants, separating agents, coating agents, sweeteners, flavoring and aroma agents, preservatives and antioxidants may also be present in the composition.

[0044] Examples of pharmaceutically usable antioxidants include: water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, and sodium sulfite; oil-soluble antioxidants such as palmitic acid ascorbate, butylated benzoic acid (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, and α-tocopherol; and metal compounds such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid.

[0045] Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions (e.g., NaCl), alcohols, gum arabic, vegetable oils, benzyl alcohol, polyethylene glycol, gelatin, sugars (e.g., lactose, amylose, or starch), polyethylene glycol, magnesium stearate, talc, silica, viscous paraffin, aromatic oils, fatty acid esters, methylcellulose, polyvinylpyrrolidone, etc. The pharmaceutical composition may be sterilized and, if desired, mixed with adjuvants such as lubricants, preservatives, stabilizers, humectants, emulsifiers, salt buffers affecting osmotic pressure, colorants, flavoring and / or aromatic substances, etc., which react harmlessly with the active compound.

[0046] The composition may also contain minor amounts of a wetting agent, emulsifier, or pH buffer. The composition may be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained-release formulation, or powder. The composition may be formulated into suppositories with conventional binders and carriers (such as triglycerides). Oral formulations may include standard carriers such as pharmaceutical-grade mannitol, lactose, magnesium stearate, polyvinylpyrrolidone, sodium saccharin, cellulose, magnesium carbonate, etc.

[0047] The composition can be formulated into a pharmaceutical composition suitable for intravenous administration to humans using conventional methods. Where necessary, the composition may also include stabilizers and local anesthetics to reduce pain at the injection site.

[0048] Generally, the ingredients are supplied individually or mixed together in unit dosage forms, such as in sealed containers like ampoules or small capsules indicating the amount of active agent as a dried, frozen powder or anhydrous concentrate. When the composition is to be administered by infusion, it can be dispersed in an infusion bottle containing pharmaceutical-grade sterile water, saline, or glucose solution. When the composition is to be administered by injection, an ampoule of sterile water or saline for injection can be provided, allowing the ingredients to be mixed prior to administration.

[0049] The pharmaceutical compositions of the present invention may further include agents for controlling the release of the compounds of the present invention, thereby providing time-controlled or sustained-release compositions.

[0050] The pharmaceutical compositions of the present invention include compositions suitable for oral, rectal, topical, vaginal, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any particular case depends on the specific subject, the nature and severity of the disease to which the active ingredient is administered. The pharmaceutical compositions can be prepared by any method known in the field of pharmaceutical science.

[0051] The active ingredient can be administered orally in solid or liquid dosage forms, such as capsules, tablets, lozenges, sugar lozenges, granules, and powders, and in liquid forms such as elixirs, syrups, emulsions, dispersions, and suspensions. The active ingredient can also be administered parenterally in sterile liquid dosage forms such as dispersions, suspensions, or solutions. Other dosage forms that can be used to administer the active ingredient include ointments, creams, drops, transdermal patches, or powders for topical administration; ophthalmic solutions or suspensions for ocular administration, i.e., eye drops; sprays or powder compositions for inhalation or intranasal administration; or creams, ointments, sprays, or suppositories for rectal or vaginal administration. Gelatin capsules contain the active ingredient and a powdered carrier, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Similar diluents can be used to prepare compressed tablets. Both tablets and capsules can be formulated as sustained-release products to provide sustained release of the drug over several hours. Compressed tablets can be sugar-coated or film-coated to mask any unpleasant tastes and protect the tablet from air, or they can be enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration may contain colorants and flavoring agents to increase patient acceptability. Generally, water, suitable oils, saline, aqueous solutions of dextran (glucose), and related sugar solutions, as well as glycols such as propylene glycol or polyethylene glycol, are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain water-soluble salts of the active ingredient, suitable stabilizers, and buffers as needed. Antioxidants such as sodium bisulfite, sodium sulfite, or ascorbic acid, alone or in combination, are suitable stabilizers. Citric acid and its salts, and sodium EDTA may also be used. In addition, parenteral solutions may contain preservatives such as benzalkonium chloride, methylparaben, or propylparaben, and chlorobutanol.

[0052] For inhalation administration, the compounds of the present invention can be conveniently delivered in aerosol form from pressurized packaging or nebulizers. The compounds can also be delivered in powder form, which can be inhaled with the aid of a powder inhaler device. A preferred delivery system for inhalation is a metered-dose inhaler (MDI) aerosol, which can be formulated as a suspension or solution of the compound of Formula I in a suitable propellant, such as a fluorocarbon or hydrocarbon. For ophthalmic administration, ophthalmic formulations can be formulated with a solution or suspension of the compound of Formula I in a suitable ophthalmic carrier at a suitable weight percentage, thereby maintaining sufficient contact time between the compound and the ocular surface to allow the compound to penetrate into the cornea and internal regions of the eye.

[0053] Useful pharmaceutical dosage forms for administering the pharmaceutical compositions of the present invention include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injections, and oral suspensions.

[0054] Fifthly, the present invention provides a method for treating a neurological disease, comprising administering to an individual an effective amount of the above-described ribramycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or a combination of the above-described pharmaceutical ingredients. This method can be used in vivo or in vitro. The individual can be a mammal, such as a human.

[0055] When the compounds of the present invention are administered stepwise or in combination with other therapeutic agents, the same dosage forms as described above may be used. When the drugs are administered in physical combinations, the dosage form and route of administration should be selected based on the compatibility of the combined drugs. The compounds of the present invention may be administered as the sole active ingredient or in combination with a second active ingredient, which includes active ingredients known to be effective in treating the relevant disease.

[0056] In some embodiments, the amount of the compound is in the range of about 0.001 mg / kg body weight / day to about 1000 mg / kg body weight / day. In other embodiments, the amount of the compound is in the range of about 0.5 mg / kg body weight / day to about 50 mg / kg body weight / day. In some embodiments, the amount of the compound is about 0.001 g / day to about 7 g / day. In other embodiments, the amount of the compound is about 0.002 g / day to about 6 g / day. In other embodiments, the amount of the compound is about 0.005 g / day to about 5 g / day. In other embodiments, the amount of the compound is about 0.01 g / day to about 5 g / day. In other embodiments, the amount of the compound is about 0.02 g / day to about 5 g / day. In other embodiments, the amount of the compound is about 0.05 g / day to about 2.5 g / day. In other embodiments, the amount of the compound is about 0.1 g / day to about 1 g / day. In other embodiments, dose levels below the lower limit of the above range may be sufficient. In other embodiments, dose levels above the upper limit of the above range may be required. In some embodiments, the compound is administered as a single dose once daily. In other embodiments, the compound is administered in multiple doses more than once daily. In some embodiments, the compound is administered twice daily. In other embodiments, the compound is administered three times daily. In other embodiments, the compound is administered four times daily. In other embodiments, the compound is administered more than four times daily. In some embodiments, the pharmaceutical composition is administered to a mammal. In other embodiments, the mammal is a human.

[0057] Preferably, the neurological disease is selected from at least one of epilepsy, Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS).

[0058] Further preferred are at least one of epilepsy and Parkinson's disease.

[0059] Terminology Explanation:

[0060] Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms, the definition provided in this chapter shall prevail.

[0061] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specified. It should also be noted that, unless otherwise specified, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0062] Definitions of standard chemical terms can be found in the references (including Carey and Sundberg, "Advanced Organic Chemistry 4th Edition," Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise stated, conventional methods within the scope of the art, such as mass spectrometry, NMR, IR, UV / Vis spectroscopy, and pharmacological methods, are used. Unless specifically defined, the terminology used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and pharmaceutical and medicinal chemistry is known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and in the treatment of patients. For example, reactions and purifications can be carried out using the manufacturer's instructions for use of kits, or in accordance with methods known in the art or the description of this invention. The techniques and methods described above can generally be carried out according to conventional methods well known in the art, based on the descriptions in the various summary and more specific references cited and discussed in this specification. In this specification, groups and their substituents can be selected by those skilled in the art to provide stable structural moieties and compounds.

[0063] When a substituent is described using a conventional chemical formula written from left to right, it also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, CH2O is equivalent to OCH2.

[0064] Unless otherwise stated, the commonly used chemical terms, such as, but not limited to, “alkyl,” “amine,” “aryl,” are equivalent to their optionally substituted forms. For example, “alkyl” as used herein includes optionally substituted alkyl groups.

[0065] The terms “optional / arbitrary” or “optionally / arbitrarily” mean that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation. For example, as defined below, “substituted or unsubstituted” means “unsubstituted” (not substituted by a substituent) or “substituted alkyl” (substituted by a substituent).

[0066] The C used in this article 1-n Including C 1-2 C 1-3 ...C 1-n For example, the "C1-C5" group refers to a portion containing 1-5 carbon atoms, meaning the group contains 1, 2, 3, or 4 carbon atoms. Therefore, for example, "C..." 1-4 "Alkyl" refers to an alkyl group having 1 to 4 carbon atoms, wherein the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Numerical ranges in this document, such as "1-10", refer to integers within a given range; for example, "1-10 carbon atoms" means that the group may have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms.

[0067] The term "alkyl" as used alone or in combination herein refers to a saturated aliphatic hydrocarbon that is optionally substituted with a straight chain or optionally substituted with a branched chain. "alkyl" as used herein preferably has 1 to about 20 carbon atoms, for example, 1 to about 10 carbon atoms, 1 to about 8 carbon atoms, 1 to about 6 carbon atoms, 1 to about 4 carbon atoms, or 1 to about 3 carbon atoms. The alkyl examples described herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, and hexyl, as well as longer alkyl groups such as heptyl and octyl. When the term "alkyl" is used in a numerical range, as defined herein, such as "C1-C6 alkyl" or "C 1-6 "Alkyl" refers to an alkyl group that can be composed of 1, 2, 3, 4, 5, or 6 carbon atoms. The term "alkyl" in this document also includes cases where no numerical range is specified.

[0068] The term “alkyl” as used in this article includes alkyl groups linked to other groups, such as alkyl in alkoxy, alkyl in alkylthio, hydroxyalkyl, haloalkyl, cyanoalkyl, monoalkylamino, dialkylamino, etc.

[0069] The term "alkoxy" as used alone or in combination herein refers to an alkyl ether group (O-alkyl), and non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

[0070] The term "aromatic / aryl" as used alone or in combination herein refers to an optionally substituted aromatic hydrocarbon group having 6 to about 20, such as 6 to 12 or 6 to 10 cyclic carbon atoms, which can be monocyclic, bicyclic, or more cyclic aryl groups. A bicyclic or more cyclic aryl group can be a monocyclic aryl group fused with other independent rings, such as alicyclic, heterocyclic, aromatic, or aromatic-heterocyclic rings. Non-limiting examples of monocyclic aryl groups include monocyclic aryl groups with 6 to about 12, 6 to about 10, or 6 to about 8 cyclic carbon atoms, such as phenyl; bicyclic aryl groups, such as naphthyl; and polycyclic aryl groups, such as phenanthryl, anthracene, or azulel.

[0071] The term "heteroaryl" as used alone or in combination herein refers to an arbitrarily substituted heteroaryl group comprising about 5 to about 20, such as 5 to 12 or 5 to 10, skeletal cyclic atoms, wherein at least one (e.g., 1-4, 1-3, 1-2) of the cyclic atoms is a heteroatom, which is independently selected from, but not limited to, heteroatoms of oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin. The ring of the group does not contain two adjacent O or S atoms. Heteroaryl groups include monocyclic heteroaryl groups (having one ring), bicyclic heteroaryl groups (having two rings), or polycyclic heteroaryl groups (having more than two rings). In embodiments where two or more heteroatoms appear in the ring, the two or more heteroatoms may be identical to each other, or some or all of the two or more heteroatoms may be different from each other. Bicyclic or more cyclic heteroaryl groups may be a monocyclic heteroaryl group fused with other independent rings, such as alicyclic, heterocyclic, aromatic, or aromatic-heterocyclic (collectively referred to as fused cyclic heteroaryl groups). Non-limiting examples of monocyclic heteroaryl groups include monocyclic heteroaryl groups with 5 to about 12, 5 to about 10, 5 to about 7 or 6 cyclic atoms in the skeleton, for example, non-limiting examples include pyridyl; fused cyclic heteroaryl groups include benzimidazolyl, quinolinyl, acridinyl. Other examples of heteroaryl compounds include, but are not limited to: pyridine, pyrimidine, pyrazine, pyridazine, triazine, furan, thiophene, imidazole, triazole, tetrazolium, thiazole, isothiazole, 1,2,4-thiadiazole, pyrrole, pyrazole, oxazole, isoxazole, oxadiazole, benzofuran, benzothiophene, benzothiazole, indole, indazole, quinoline, isoquinoline, purine, carbazole, benzimidazole, pyrrolopyridine, pyrrolopyrimidine, pyrazolopyridine, pyrazolopyrimidine, etc.; acridine, phenazinyl, benzoxazolyl, benzothiadiazoleyl, benzoxadiazoleyl, benzotriazoleyl, isoquinolinyl, indolizinyl, isothiazolyl ( isothiazolyl), isoindolyl, oxadiazolyl, purinyl, phthalazinyl, pteridinyl, quinazolinyl, quinoxalinyl, triazinyl, and thiadiazolyl, and their oxides, such as pyridyl-N-oxide.

[0072] The term "heterocycle" or "heterocyclic group" as used alone or in combination herein refers to a non-aromatic heterocycle, including saturated heterocycles or unsaturated heterocycles (containing unsaturated bonds). One or more of the cyclic atoms (e.g., 1-4, 1-3, 1-2) are heteroatoms, such as oxygen, nitrogen, or sulfur atoms. Heterocycles can include monocyclic (having one ring), bicyclic (having two bridging rings), or polycyclic (having more than two bridging rings); spirocyclics are also included. Heterocyclic groups can have from 3 to about 20, such as 3-about 10, 3-about 8, 4-8, 4-7, 5-about 8, or 5-about 6 cyclic atoms. Non-limiting examples of heterocyclic groups include azinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, and pyrazolinyl. The term also includes cyclohexyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl, and quinolizinyl, among others. The term also includes all cyclic forms of sugars, including but not limited to monosaccharides, disaccharides, and oligosaccharides. Examples also include, but are not limited to, aziridine, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, oxazolidine, thiazolidinidine, imidazolidinidine, isoxazolidine, isothiazolidine, pyrazolidine, morpholine, thiomorpholine, piperazine, piperidinyl, etc.Heterocyclic groups also include heterocycles with one or more fused aromatic rings (i.e., sharing a common bond), such as 2,3-dihydrobenzofuran, 1,3-benzodioxane, benzo-1,4-dioxane, phthalimide, and naphthalenedioximide. Heterocyclic groups with one or more fused aromatic rings can be linked to other groups through the aromatic or non-aromatic ring portions. It should be noted that heterocyclic groups include substituted heterocyclic groups, meaning that other groups can be linked to the heterocycle through heteroatoms or carbon atoms (i.e., the heterocycle is linked to the parent molecule or further substituted).

[0073] "Halogen" refers to fluorine, chlorine, bromine, and iodine. Fluorine, chlorine, and bromine are preferred. A cyano group is "-CN"; a hydroxyl group is "-OH"; a mercapto group is "-SH"; and an amino group is "-NH2".

[0074] The term "ester group" has two linkage forms, including R1-OCO-R2 and R1-COO-R2, where R1 and R2 refer to the substituents on both sides, and R1 and R2 can be the same or different.

[0075] The term "substituted" means that one or more hydrogen atoms on a particular atom are replaced by a specified group. If the normal valence of the specified atom is not exceeded under the existing conditions, then the result of the substitution is a stable compound.

[0076] Unless otherwise stated, all ranges listed in this article are inclusive. For example, "n is an integer between 0 and 2" means that n can be 0, 1, or 2.

[0077] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable, non-toxic alkali or acid. When the compounds of the present invention are acidic, their corresponding salts can be readily prepared from inorganic or organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganese), potassium, sodium, zinc, etc. Preferred are ammonium, calcium, magnesium, potassium, and sodium salts. Salts prepared from organic bases include primary, secondary, and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable non-toxic organic bases that can form salts include arginine, betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, dicyclohexylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compounds of the present invention are basic, their corresponding salts can be readily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, hydroxyethylsulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, viscous acid, nitric acid, pyric acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc.

[0078] The term "solvate" refers to a variable stoichiometric complex formed by a solute (i.e., a compound of formula I) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is called a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, sesquihydrates, dihydrates, and trihydrates.

[0079] The term "prodrug" refers to a functional derivative of the compound of the present invention that is readily converted into the desired compound in vivo. Various forms of prodrugs are well known in the art. See also the discussion of prodrugs in T. Higuchi and V. Stella's *Pro-drugs as Novel Delivery Systems* (1987), Vol. 14 of the ACSSymposium Series, *Bioreversible Carriers in Drug Design* (1987), Edward B. Roche, ed., American Pharmaceutical Association, and Pergamon Press. Design of Prodrugs, Bundgaard, A. Ed., Elseview, 1985 and Method in Enzymology, Widder, K. et al., Ed.; Academic, 1985, vol. 42, pp. 309-396; Bundgaard, H. "Design and Application of Prodrugs" in A Textbook of Drug Design and Development, Krosgaard-Larsen and H. Bundgaard, Ed., 1991, Chapter 5, pp. 113-191; and Bundgaard, H., Advanced Drug Delivery Review, 1992, 8, 1-38. These references are incorporated herein by reference.

[0080] "Subject," "patient," or "individual" refers to an individual suffering from a disease, symptom, or condition, including both mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans; non-human primates (e.g., chimpanzees and other apes and monkeys); livestock such as cattle, horses, sheep, goats, and pigs; domesticated animals such as rabbits, dogs, and cats; and laboratory animals, including rodents such as rats, mice, and guinea pigs. Examples of non-human mammals include, but are not limited to, birds and fish. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0081] The terms "treatment and / or prevention" and other similar synonyms include relieving, reducing, or improving symptoms of a disease or condition; preventing other symptoms; improving or preventing the underlying metabolic causes of symptoms; inhibiting a disease or condition, such as preventing its progression; alleviating a disease or condition; improving a disease or condition; relieving symptoms caused by a disease or condition; or stopping the symptoms of a disease or condition. Furthermore, the term includes the purpose of prevention. The term also includes achieving therapeutic and / or preventive effects. A therapeutic effect refers to the cure or improvement of the underlying disease being treated. Additionally, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect; for example, an improvement in the patient's condition is observed even though the patient may still be affected by the underlying disease. In terms of preventive effects, the composition may be administered to patients at risk of developing a specific disease, or to patients exhibiting one or more physiological symptoms of a disease, even if no disease diagnosis has been made.

[0082] As used herein, the terms "effective amount," "therapeutic effective amount," or "pharmaceutical effective amount" refer to an amount of at least one drug or compound that, when taken, is sufficient to alleviate, to some extent, one or more symptoms of the disease or condition being treated. The result may be a reduction and / or relief of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition containing the compounds disclosed herein that is clinically necessary to provide significant symptom relief. Effective amounts suitable for any individual case can be determined using techniques such as dose escalation testing.

[0083] As used herein, the terms “administration,” “application,” “dosage,” etc., refer to methods that deliver a compound or composition to the desired site for biological action. These methods include, but are not limited to, oral administration, duodenal administration, parenteral administration (including intravenous, subcutaneous, intraperitoneal, intramuscular, intra-arterial injection or infusion), topical application, and rectal administration. Those skilled in the art are familiar with administration techniques that can be used with the compounds and methods described herein, such as those discussed in Goodman and Gilman, *The Pharmacological Basis of Therapeutics*, current ed.; Pergamon; and Remington's, *Pharmaceutical Sciences* (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

[0084] The term "acceptable" as used in this article for formulations, compositions or ingredients means that it has no long-term harmful effects on the general health of the subject receiving the treatment.

[0085] As used herein, the term "pharmaceutically acceptable" means a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, i.e., that the substance can be administered to an individual without causing an adverse biological reaction or interacting adversely with any component contained in the composition.

[0086] The term "pharmaceutical composition" refers to a bioactive compound that optionally contains at least one pharmaceutically acceptable chemical component, including but not limited to carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients.

[0087] The term "carrier" refers to a relatively non-toxic chemical compound or reagent that helps to introduce the compound into cells or tissues.

[0088] The term "nervous system disease" refers to diseases that occur in the central nervous system, peripheral nervous system, and autonomic nervous system, and are characterized by sensory, motor, consciousness, and autonomic nervous system dysfunction.

[0089] The beneficial effects of this invention are as follows: This invention provides a ribramycin derivative and its use in neurological diseases, expanding the application of ribramycin derivatives. At the same time, for neurological diseases of the brain, this invention designs a compound with a new structure. This compound has excellent brain-passing performance, overcoming the current problem of low efficiency of drug passage through the brain, and showing greater advantages in the treatment of neurological diseases of the brain. Attached Figure Description

[0090] Figure 1 shows the mass spectrometry detection of compound I of the present invention.

[0091] Figure 2 shows the mass spectrometry of compound I-2 of the present invention.

[0092] Figure 3 shows the mass spectrometry of compound I-3 of the present invention.

[0093] Figure 4 shows the mass spectrometry of compound I-4 of the present invention.

[0094] Figure 5 shows the statistical analysis of IC50 in HepG2 cells after treatment with different concentrations of 18-Me-Reblastatin and 17AAG.

[0095] Figure 6 shows the cytotoxicity results of I-2, I-3, and I-4.

[0096] Figure 7 shows the experimental results of increasing GLT1 protein levels in primary cultured astrocytes.

[0097] Figure 8 shows the GLT1 detection results. The left side of the figure shows the Western blotting analysis results of mouse hippocampal tissue homogenate after 18-Me-Reblastatin injection, which detected the target protein GLT1 and the internal reference protein Actin, respectively. The right side shows the statistical analysis results of the relative changes in GLT1 protein levels.

[0098] Figure 9 shows the effect of PTZ-induced acute epilepsy and the protective effect of 18-Me-reblastatin, with data combined from two animal experiments.

[0099] Figure 10 shows the statistical results of the frequency of epileptic seizures in male APP / PS1 mice.

[0100] Figure 11 shows the results of the mouse pole climbing and turning tests. Detailed Implementation

[0101] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the invention based on the disclosed content, which should also fall within the scope of protection claimed in this application.

[0102] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention were obtained through conventional commercial means. Unless otherwise specified, all contents mentioned below are mass contents. Unless otherwise specified, it is understood that the process was carried out at room temperature.

[0103] The compounds of the present invention having the various forms described herein can be prepared from commercially available starting materials or starting materials that can be prepared using literature processes, according to the processes described in the following general synthetic routes. Variables (e.g., R) in each general synthetic route are as defined herein. Those skilled in the art will note that the order of certain steps in the reaction procedures and synthetic schemes described herein can be varied, such as the introduction and removal of protecting groups.

[0104] In this invention, 18-Me-Reb and 18-Me-Reblastatin both refer to compound I.

[0105] Compound preparation

[0106] Example 1: Preparation of 18-Me-Reblastatin (Compound I)

[0107] (1) Preparation of 18-OTf-Reblastatin (compound III)

[0108] Compound II (274.3 mg) was dissolved in anhydrous DMF (2 mL), and DIPEA (0.26 mL), Tf2NPh (268 mg), and DMAP (7 mg) were added. The reaction flask was then heated in an oil bath at 40 °C for 5 hours. TLC monitoring showed that the starting material spot disappeared. The reaction solution was cooled to room temperature, the reaction was quenched with water, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to give compound III (301 mg, yield 88%).

[0109] ESI (m / z): 703.72 [M+Na] +

[0110] (2) Preparation of 18-Me-Reblastatin (compound I)

[0111] Compound III (301 mg), K3PO4 (280 mg), and Pd(PPh3)4 (31 mg) were added to a single-necked round-bottom flask. The mixture was then protected with argon gas, followed by the addition of dioxane (10 mL). The mixture was degassed by sonication, and then 151 μL of TMB in THF solution was added. The reaction flask was then heated in an oil bath (100 °C) for 4 hours. After the reaction was complete as monitored by TLC, the reaction mixture was cooled to room temperature, quenched with water, extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain the target compound I (200 mg, yield 83%).

[0112] ESI (m / z): 569.32 [M+Na] +

[0113] Example 2 Preparation of compound I-2

[0114] Compound III (14 mg), KOAc (8 mg), and Pd(dppf)2Cl2 (2 mg) were added to a round-bottom flask, and argon protection was applied. Dioxane (1 mL) was added, and the mixture was degassed by sonication. The reaction solution was then heated in an oil bath (100 °C) for 1 hour. The reaction solution was cooled to room temperature, diluted with ethyl acetate, and the reaction was quenched with water. The mixture was separated, extracted with ethyl acetate, and the organic phases were combined and washed with saturated brine. The mixture was dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain the target compound I-2 (7 mg, yield 52%).

[0115] ESI (m / z): 681.39 [M+Na] +

[0116] Example 3 Preparation of compound I-3

[0117] Compound I-3 (20 mg) was added to a round-bottom flask, and the mixture was protected with argon gas. DMF (2 mL) was added, and the reaction solution was then cooled in an ice-water bath. Potassium carbonate (10 mg) and acyl chloride (5 μL) were added, and the mixture was then heated to room temperature and reacted for 1 hour. After the reaction was complete, the mixture was diluted with ethyl acetate, and water was added for liquefaction. The ethyl acetate was used for extraction, and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain the target compound I-3 (8 mg, yield 35%). A portion of the starting material (10 mg, recovery 50%) was also recovered.

[0118] ESI (m / z): 620.20 [M+Na] +

[0119] Example 4 Preparation of compound I-4

[0120] Compound I-3 (5 mg) was added to a round-bottom flask, and the mixture was protected with argon gas. DMF (0.5 mL) was added, and the reaction solution was then cooled in an ice-water bath. Potassium carbonate (3 mg) and acyl chloride (2 mg) were added, and the mixture was then brought to room temperature and reacted for 1 hour. After the reaction was complete, the mixture was diluted with ethyl acetate, diluted with water, extracted with ethyl acetate, and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain the target compound I-4 (4 mg, 65% yield).

[0121] ESI (m / z): 698.23 [M+Na] +

[0122] Effect detection

[0123] 1. Cytotoxicity detection

[0124] We used HepG2 cells to detect cell proliferation inhibition rate using CTG (CELL TITER-GLO) luminescence assay. We screened several compounds at concentrations of 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20, and 50 μM, respectively, and cultured them for 2 days in a 37℃, 5% CO2 incubator before CTG detection. The IC50 was calculated, and the results are shown in Figure 5. It can be seen that the IC50 of 17AAG is 0.017 μM, and the toxicity of 18-Me-Reb is 0.349 μM, which is much lower than the toxicity of 17AAG (19.5 times).

[0125] Meanwhile, we also evaluated the cytotoxicity of I-2, I-3, and I-4, as shown in Figure 6. We found that the IC50 values ​​were 0.301 μM, 0.238 μM, and 0.347 μM, respectively, which were similar to those of 18-Me-Reb and were all much lower than the toxicity of 17AAG.

[0126] 2. Experiment to increase GLT1 protein levels in primary cultured astrocytes

[0127] To investigate whether the four compounds could increase GLT1 expression levels, we cultured primary astrocytes and treated them with the four compounds on day 14 post-culture. The treatment doses were 0, 20, 50, 100, 200, and 400 nM. Forty-eight hours after treatment, astrocytes were collected for Western blotting to detect GLT1 and Actin protein levels. The results are shown in Figure 7, indicating that all four compounds increased GLT1 protein levels.

[0128] 2. Experiment on the effect of 18-Me-reblastatin on increasing GLT1 protein levels in mouse hippocampal brain tissue

[0129] Nine-week-old C57 / BL6j mice, 20 males and 4 females in each dose group, were used. They were administered intraperitoneally at doses of 0, 1, 2, 4, and 8 mg / kg. Hippocampal brain tissue was harvested on the morning of day 6 after injection on days 1, 3, and 5. Tissue homogenate was extracted, and 10 μL (20 μg) of protein was analyzed by Western blotting. One-way ANOVA was used for statistical analysis. As shown in Figure 8, the results indicated that GLT1 protein levels increased gradually in a dose-dependent manner, reaching a maximum at a dose of 4 mg / kg.

[0130] 3. Blood-brain barrier permeability test

[0131] 3.1 Blood-brain barrier permeability test of compound I

[0132] First, the plasma concentration of the compound was measured in mice after a single intraperitoneal injection at a dose of 4 mg / kg. Relevant pharmacokinetic parameters and the brain-blood ratio of the compound were calculated. The time to peak plasma concentration was 0.14 hours, and the half-life (T1 / 2) was 2.07 hours. By measuring the plasma and brain concentrations of the compound at 0.083, 0.167, and 0.5 hours post-injection, the brain-blood ratios at these three time points were 0.35, 0.52, and 0.54, respectively. This indicates that approximately half of the compound can pass through the brain, achieving the properties of a central nervous system drug.

[0133] Table 1. Blood drug concentration at different time points after a single intraperitoneal injection of 18-Me-Reblastatin

[0134] In Table 3-8, Mean: mean, SD: standard deviation, CV: coefficient of variation, BLQ: below the limit of quantitation (not detected), NA: unavailable.

[0135] Table 2. Pharmacokinetic parameters of 18-Me-Reblastatin

[0136] In Table 2, IP: intraperitoneal injection, Mean: mean, SD: standard deviation, HL-Lambda-Z: half-life, and AUC: area under the concentration-time curve, representing the bioavailability of the drug (the degree to which the drug is absorbed and utilized in the human body). A larger AUC indicates higher bioavailability, and vice versa. (0-t) AUC from 0 to the final quantifiable time point, AUC (0-∞) AUC, T from 0 to infinity max Time to peak drug concentration, C max Peak drug concentration; AUClast: AUC from the start of administration to the last point; AUCINF-obs: AUC from the start of administration to theoretical extrapolation infinity; MRTlast: Mean residence time from the start of administration to the final quantifiable concentration; MRTINF-obs: Mean residence time of the drug from the start of administration to theoretical extrapolation infinity; T 1 / 2 Drug half-life, MRT (0-t) MRT (Mean Time Retention) refers to the average residence time of the drug from 0:00 to the final quantifiable time point. (0-∞) : The average residence time of the drug from 0 to infinity.

[0137] Table 3. Brain concentration detection 0.083 hours after a single intraperitoneal injection of 18-Me-Reblastatin

[0138] Table 4. Cerebral-to-blood ratio 0.083 hours after a single intraperitoneal injection of 18-Me-Reblastatin

[0139] Table 5. Brain concentration detection 0.167 hours after a single intraperitoneal injection of 18-Me-Reblastatin

[0140] Table 6. Cerebral-to-blood ratio 0.167 hours after a single intraperitoneal injection of 18-Me-Reblastatin

[0141] Table 7. Brain concentration detection 0.5 hours after a single intraperitoneal injection of 18-Me-Reblastatin

[0142] Table 8. Cerebral blood-to-reblastatin ratio 0.5 hours after a single intraperitoneal injection of 18-Me-Reblastatin

[0143] In Table 3-8, Mean: mean, SD: standard deviation, and CV: coefficient of variation.

[0144] 3.2 Blood-brain barrier permeability test of Reblastatin

[0145] The plasma and brain tissue concentrations of Reblastatin in mice after intraperitoneal injection were measured using the method described above. It was found that Reblastatin was almost undetectable in the brain. No concentration of the compound was detected in the brain 0.5 hours after intraperitoneal injection, indicating that its brain-passing efficiency was low.

[0146] Table 9. Detection of Reblastatin blood and brain tissue concentrations

[0147] In Table 9, IP: intraperitoneal injection, Mean: mean, SD: standard deviation, CV: coefficient of variation, BLQ: below the limit of quantitation (not detected), NA: unavailable.

[0148] 4. Protective effect of compound I in pentylenetetrazol (PTZ)-induced acute epilepsy.

[0149] PTZ is a central nervous system stimulant that primarily induces seizures by acting as a non-competitive antagonist of GABA receptors. This inhibition reduces GABA-mediated inhibitory neurotransmission, leading to an imbalance between excitatory and inhibitory signals in the brain. Consequently, increased neuronal excitability triggers acute epileptic seizures. PTZ-induced acute epilepsy models are frequently used for the rapid screening of antiepileptic and anti-neurostimulatory compounds.

[0150] Nine-week-old male C57 / BL6j mice were used. There were 18 mice in the control group and 15 mice in the 18-Me-Reblastatin group. On days 1, 3, and 5, mice were intraperitoneally injected with 4 mg / kg of the compound or solvent dissolved in DMSO (100 μL). On day 6 at noon, PTZ (potassium phosphate) was injected intraperitoneally at a dose of 55 mg / kg to induce seizures. The time of onset of the first seizure after PTZ injection and the Racine scale score of the highest-grade seizure were recorded.

[0151] Racine rating criteria:

[0152] 0) No seizure behavior;

[0153] 1) The behavior suddenly stops, and the person stares motionless;

[0154] 2) Beard trembling and / or facial and neck twitching;

[0155] 3) Seated clonic seizures;

[0156] 4) Tonic-clonic seizures (while lying prone);

[0157] 5) Tonic-clonic seizures (lying on one's side, losing postural control) or violent jumping.

[0158] The statistical method used was Student's t-test, and the results are shown in Figure 9. The results show that the average seizure onset time in the solvent control group was 80 seconds, while in the 18-Me-Reblastatin treatment group it was 98 seconds. Regarding the Racine score, the average score in the solvent control group was 3.8, while the average score in the 18-Me-Reblastatin treatment group was 2.9. In conclusion, 18-Me-Reblastatin pretreatment prolonged the onset time of PTZ-induced seizures and reduced their severity. This indicates that 18-Me-Reblastatin has a protective effect against PTZ-induced acute epilepsy.

[0159] 5. The anti-epileptic and cognitive function-improving effects of compound I in an AD mouse model (APP / PS1)

[0160] Eight-month-old male APP / PS1 mice (carrying both APP695swe and PS1-dE9 mutations; Jacksonlab strain number 034832) were used. Eight mice were assigned to each of the control and experimental groups for electroencephalography (EEG) monitoring. After electrode implantation, baseline EEG recordings were continuously monitored for 24 hours × 14 days. Starting on day 15, either the solvent control group or 18-Me-Reblastatin (4 mg / kg) was administered intraperitoneally every other day for seven consecutive days. EEG recordings were discontinued on day 14 after the first injection. The effects of the compounds were analyzed by comparing baseline and post-treatment seizure frequency. As shown in Figure 10, the results showed that the solvent (DMSO) control group had no significant effect on seizure frequency, with a baseline frequency of 1.08 seizures / day and a post-solvent injection frequency of 1.11 seizures / day. In the experimental group, the baseline seizure frequency was 0.96 seizures / day, which decreased to 0.16 seizures / day after 18-Me-Reblastatin treatment, representing an average decrease of 83% in seizure frequency.

[0161] 6. Protective effect of compound I in MPTP-induced PD model

[0162] To evaluate the therapeutic effect on a mouse model of Parkinson's disease, 37 eight-week-old male c57 mice were randomly divided into three groups: a control group (n=8), an MPTP modeling group (n=11), and an 18Me treatment group (n=18). The treatment groups were pre-administered with 18-Me-Reblastatin (4 mg / kg, ip) twice, followed by MPTP subacute PD modeling. MPTP was administered intraperitoneally at a dose of 30 mg / kg to the mice, while the control group received an equal volume of saline, once daily for 14 days. During this period, the treatment groups were simultaneously intraperitoneally injected with 18-Me-Reblastatin (4 mg / kg) every other day, while the control and modeling groups received an equal volume of DMSO. Behavioral assessments were performed two weeks later.

[0163] Test 1: Pole Climbing Test. This test assesses the motor coordination of mice by observing their ability to grasp and descend a vertical pole. The experimental setup consists of a 1 cm diameter, 50 cm long wooden pole with a 2.5 cm diameter wooden ball fixed to the top. The pole is wrapped in gauze to prevent slipping. During the test, the pole is at a 90° angle to the ground. The mouse is placed on the top ball, and the time taken for the mouse to spontaneously climb from the top to the bottom is recorded. Each mouse is tested three times, with at least a 30-minute interval between tests, and the average of the three results is taken.

[0164] Test 2: Roulette test. Used to assess the coordination and balance of mice. Each mouse was pre-trained for three days before all tests, with speeds set at 8 rpm, 12 rpm, and 16 rpm for 5 minutes each. During the actual test, the roulette wheel was set to a uniform acceleration from 4 rpm to 40 rpm for 10 minutes, and the time it took for the mouse to fall from the rotating bar was recorded. Each mouse was tested three times, with at least 30 minutes between each test, and the average of the three results was taken.

[0165] The results, shown in Figure 11, indicate that 18-Me-Reblastatin can significantly alleviate motor damage caused by MPTP. This suggests that 18-Me-Reblastatin also has a protective effect against PD.

[0166] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. Use of a ribramycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs in the preparation of a treatment for and / or prevention of neurological disorders. The structural formula of the riburamycin derivative is shown in Formula I: in, R is selected from H; halogen; amino; substituted or unsubstituted C1-5 alkyl, C1-5 alkoxy, C3-10 cycloalkyl, C1-5 ester, aryl, amino, heterocyclic, wherein the substituted group is selected from at least one of C1-C5 alkyl, carbonyl, hydroxyl, C1-C5 alkyl, halogen, amino, aryl, heterocyclic.

2. The use according to claim 1, characterized in that, The R is selected from substituted or unsubstituted C1-5 alkyl, C1-5 alkoxy, C3-10 cycloalkyl, ester, aryl, amino, heterocyclic groups, and the substituted group is selected from at least one of C1-C5 alkyl, carbonyl, hydroxyl, C1-C5 alkyl, halogen, amino, aryl, and heterocyclic groups.

3. The use according to claim 2, characterized in that, The R is selected from substituted or unsubstituted C1-5 alkyl, C1-5 ester, or heterocyclic groups. The substituted group is selected from at least one of carbonyl, hydroxyl, C1-C5 alkyl, halogen, amino, aryl, or heterocyclic groups. The heteroatom in the heterocyclic group is selected from at least one of N, S, B, or O.

4. The use according to claim 3, characterized in that, The R is selected from substituted or unsubstituted C1-3 alkyl, C1-5 ester, or heterocyclic groups, and the substituted group is selected from at least one of C1-C5 alkyl, amino, or heterocyclic groups; the heteroatom in the heterocyclic group is selected from at least one of N, S, B, or O.

5. The use according to claim 4, characterized in that, The R is selected from C1-3 alkyl, -OCO-Ra, substituted heterocyclic groups, and the Ra and the substituted substituents are each independently selected from at least one of C1-C3 alkyl, amino, and heterocyclic groups; the heteroatom in the heterocyclic group is selected from at least one of N, S, B, and O.

6. The use according to claim 5, characterized in that, The R is selected from C1-3 alkyl, -OCO-Ra, substituted heterocyclic groups, and the Ra is selected from at least one amino group or heterocyclic group; the substituted substituent is selected from C1-C3 alkyl, and the heteroatom in the heterocyclic group is selected from at least one of N, B, and O.

7. The use according to claim 6, characterized in that, The R is selected from C1-3 alkyl, -OCO-Ra, or at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B or O, and the Ra is selected from at least one amino group or N-containing six-membered heterocyclic group.

8. The use according to claim 7, characterized in that, The R is selected from methyl, -OCO-Ra, at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B and O, and the Ra is selected from at least one C1-3 alkyl-substituted amino group, piperazine group, and C1-3 alkyl-substituted piperazine group.

9. The use according to claim 8, characterized in that, The R is selected from methyl, -OCO-Ra, at least one methyl-substituted pentacyclic heterocyclic group containing B or O, and the Ra is selected from at least one of dimethylamino, methylethylamino, diethylamino, piperazine, or methyl-substituted piperazine.

10. The use according to claim 9, characterized in that, The R is selected from methyl, -OCO-Ra, Ra is selected from dimethylamino, 11. The use according to claim 10, characterized in that, R is a methyl group.

12. The use according to any one of claims 1-11, characterized in that, The neurological disease in question is a brain disease.

13. The use according to claim 12, characterized in that, The neurological disease is selected from at least one of epilepsy, Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS).

14. The use according to claim 13, characterized in that, The neurological disease is selected from at least one of epilepsy and Parkinson's disease.

15. A riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs, characterized in that, The structural formula of the riburamycin derivative is shown in Formula I: The R is selected from methyl, -OCO-Ra, or substituted heterocyclic groups, and the Ra and the substituted substituents are each independently selected from at least one of C1-C3 alkyl, amino, or heterocyclic groups; the heteroatom in the heterocyclic group is selected from at least one of N, S, B, or O.

16. The riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs according to claim 15, characterized in that, The R is selected from C1-3 alkyl, -OCO-Ra, substituted heterocyclic groups, and the Ra is selected from at least one amino group or heterocyclic group; the substituted substituent is selected from C1-C3 alkyl, and the heteroatom in the heterocyclic group is selected from at least one of N, B, and O.

17. The riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs according to claim 16, characterized in that, The R is selected from C1-3 alkyl, -OCO-Ra, or at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B or O, and the Ra is selected from at least one amino group or N-containing six-membered heterocyclic group.

18. The riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs according to claim 17, characterized in that, The R is selected from methyl, -OCO-Ra, at least one C1-3 alkyl-substituted pentacyclic heterocyclic group containing B and O, and the Ra is selected from at least one C1-3 alkyl-substituted amino group, piperazine group, and C1-3 alkyl-substituted piperazine group.

19. The riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs according to claim 18, characterized in that, The R is selected from methyl, -OCO-Ra, at least one methyl-substituted pentacyclic heterocyclic group containing B or O, and the Ra is selected from at least one of dimethylamino, methylethylamino, diethylamino, piperazine, or methyl-substituted piperazine.

20. The riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs according to claim 19, characterized in that, The R is selected from methyl, -OCO-Ra, Ra is selected from dimethylamino, 21. The riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs according to claim 20, characterized in that, R is a methyl group.

22. The method for preparing the ribramycin derivative according to any one of claims 15-21, characterized in that, Choose from the following methods: Wherein, R has the same definition as that described in any one of claims 15-21.

23. The preparation method according to claim 20, characterized in that, When R is a methyl or substituted heterocyclic group, the preparation method is as follows: When R is -OCO-Ra, the preparation method is as follows:

24. A pharmaceutical composition, characterized in that, Includes the riburamycin derivative or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs as described in any one of claims 15-21, and pharmaceutically acceptable carriers or excipients.

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